Analysis of a portrait of Queen Elizabeth I by East Carolina University researchers showed that it was from the 1590s rather than a more recent copy. (Joe Barricella/Courtesy of East Carolina University)

More than 100 men, women and children who were trying to build the first English settlement in the New World vanished from North Carolina’s Roanoke Island four centuries ago. Now the same thing has happened to a beloved if unflattering 16th-century portrait of their monarch, Queen Elizabeth I.

The rare painting of the English ruler in the last years of her reign resided for more than 60 years at the site of the “Lost Colony.” Proud locals dubbed it “the Manteo Queen” after a nearby island town.

Late last year, however, the garden club that owned the portrait shipped it quietly to Britain and sold it at Sotheby’s for $60,000, although it had a $100,000 appraisal. The buyer, Philip Mould, a London dealer in historic portraits, said he has since resold it to a private collector “enthusiastic of royalty.” “It will be staying in England,” said Mould, who stars in a BBC television show called "Fake or Fortune" and long served as art adviser to Parliament. He declined to name the selling price.

The abrupt loss of the monarch’s portrait to an overseas dealer — there are only a few of her portraits in the United States — has outraged the American scholars who helped bring international attention to the painting in 2010 after a battery of scientific tests. “I feel like I’ve been kicked in the gut,” said Larry Tise, a historian at East Carolina University in Greenville, N.C., who spearheaded the analysis of the painting. “This unique portrait — both as a work of art and as a historical artifact — was a great treasure to North Carolina.”

But for the cash-strapped Elizabethan Gardens that owned it, the sale provided a new slate roof and paint job for the organization’s gatehouse, according to executive director Carl Curnutte. The nonprofit organization manages a series of gardens adjacent to the Fort Raleigh National Historic Site on the shores of Albemarle Sound.

Until 2010, the painting hung for nearly 60 years just inside the door of that gatehouse. A North Carolina benefactor named Ruth Coltrane Cannon had purchased it from a New York gallery in 1958 and presented it as a gift to the organization. It was with the queen’s approval that her favored courtier, Sir Walter Raleigh, organized the Roanoke venture. The project ended in the loss in 1587 of the settlers, including Virginia Dare, the first English child born in the New World. Their fate has never been determined. The failed effort was followed 20 years later by the first successful English settlement, the Jamestown colony in 1607, and ultimately, the creation of the United States.

On a visit in 2008, Tise saw the portrait and decided to use modern techniques to determine whether it was an Elizabethan portrait or a later copy. Two years later, with the permission of the Elizabethan Gardens, he and colleagues from East Carolina University subjected the portrait to nondestructive tests of ultraviolet and infrared radiation, as well as X-ray fluorescence.

The team concluded that the image was not traced but drawn freehand, a common practice in the late 16th century. That meant it was more than an exact copy made by one artist from the work of another. And an underlying drawing, unseen in visible light, revealed that the image was based on specific dress and facial patterns. These were in use in the final era of Elizabeth’s reign in artist workshops that specialized in portraiture. Also, the painting was found to be made on Baltic oak, which was commonly used for Elizabethan-era royal portraits but which ceased to be imported into England by the middle of the 17th century. Based on these findings, the painting was likely produced in the 1590s.

“Our tests nailed the date,” said Sara James, an emeritus art historian at Mary Baldwin College in Virginia who was involved in the data gathering. The painting subsequently was housed in a secure storage facility owned by the National Park Service, though it was shown in 2013 at the Folger Shakespeare Library in Washington. A new insurance appraisal based on the 2010 research study put its value at $100,000.

The tests also showed that decades of exposure to the heat and humidity of coastal North Carolina had done little to damage the portrait. But there was evidence that in past centuries the portrait had been cleaned with strong solvents, fuzzing details of the sumptuous clothes, jewelry and hair, some of which were later retouched. The face, however, appears to be better preserved than other portions of the image.

And it is the face that makes the painting unusual. The visage of the Manteo Queen is deeply lined and withered. This is a remarkable deviation from most of the other hundreds of portraits made of the English ruler.

“The queen was incredibly anxious about the dissemination of her portraits,” said Edward Town, a Yale University art historian. As a result, few images show her as an old woman. In 1596, when the Virgin Queen was 63 years old, the government ordered that any portrait that caused her “great offence” should be collected and burned. As a result, there are no known paintings of the queen based on royal sittings between 1596 and her 1603 death. This portrait is probably not an exception. James said that the underdrawing suggests that the artist added wrinkles and creases to the monarch’s face without actually seeing her. Still, “the Manteo Queen still is among only a few portraits to show the ravages of time,” she said.

Who painted the image remains controversial. “The debate is whether it was a product of the workshop of Marcus Gheeraerts the Younger or done by someone with access to his patterns,” Town said.

George Perry Hurt, associate conservator at the North Carolina Museum of Art in Raleigh who was part of the analysis team, argued that it likely was done in the provinces rather than London. Mould agreed with that assessment, and said he was confident of a production date in the 1590s. “It was probably done by a provincial follower of Gheeraerts,” he added. “But it is a portrait of considerable competence.”

Mould encountered the Manteo Queen in 2013, when he commented favorably on its authenticity in the CNBC series "Treasure Detectives." Curnutte said Mould bought the portrait for less than the minimum price of $60,000 set by Sotheby’s. Sources close to the sale said he paid $51,000. When Curnutte complained, he said the auction house made up the difference. The Gardens, he added, netted $46,000 from the sale.

Mould would not name the new owner, or the price for which he flipped the painting. In 2007, Mould purchased the Hampden portrait of Elizabeth for $3.4 million. In July, the monarch’s famous Armada portrait was sold for nearly $12.5 million in London, thanks to a large grant from the British national lottery and public contributions. By contrast, a small circular bust-length portrait sold for $30,000 in October at Christie’s in New York.

“It is difficult to predict the price of monarch portraits,” said one art historian. “Demand is erratic and its value not just tied to the quality.” Added another: “If Mould turned around and sold it for half a million, that’s the art market.”

Sotheby’s is embroiled in a legal battle involving the former owner of a newly recognized work by Leonardo da Vinci. It sold for $80 million and the buyer, a billionaire and longtime Sotheby‘s client, promptly resold it for $127.5 million. The former owner intends to sue Sotheby’s, which last month filed papers in New York to block the suit.

Meanwhile, Tise and James, who wanted the unusual portrait to remain at the site of the first English settlement attempt, and who only found out about the sale months after the fact, are angry at the loss. “To me it’s tragic,” said James. “I don’t know how they” — the Elizabethan Gardens — “got so hoodwinked. They just didn’t value it.”

Curnutte defended a decision that he said was made after a prolonged debate among board members. “We are a small nonprofit without the means to take care of it, and putting it in storage didn’t seem right,” he added. “We couldn’t hang this great work of art in our gatehouse. I have no seller’s remorse.”